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DNA adsorption on silica, alumina and hydroxyapatite and imaging of dna by atomic force microscopy
The scope of the study is to investigate calf thymus DNA adsorption process on solid powders such as silica, alumina and hydroxyapatite (HAP) to improve DNA solid phase extraction efficiency and to visiulize DNA adsorption by atomic force microscopy (AFM). The equilibrium and kinetics of the DNA adsorption were investigated in batch adsorption on a commercial silica gel and a synthesized silica aerogel, commercial alumina and HAP powders from aqueous DNA solution. Commercial DNA extraction kit adsorbents were also characterized and used for adsorption. Adsorbed DNA was imaged in ambient air on flat surfaces of mica, silica and alumina wafers and HAP particles coated glass surfaces and a HAP pellet surface by AFM. Effects of ambient air, nitrogen flow and freeze drying methods on DNA morphology on the releted surfaces were also investigated. Adsorption of DNA on silica, alumina and HAP increased with the decrease of pH from 9.0 to 2.0. Enhancements of the adsorption capacities of adsorbents were obtained with the addition of MgCl2. It was found that the Van der Waals and the hydrogen bonds rather than the surface charge were the cause of the adsorption. The adsorption isotherms of DNA on silica, alumina and HAP were fitted to Langmuir model in pH range 2-9. The adsorption kinetics obeyed pseudo second order model. The sharpest image of DNA by AFM was obtained by freeze drying method on alumina surface. Dispersed DNA in water was adsorbed on the surfaces not as single molecules but as supercoils consisting of many molecules.
- Phd Degree / Doktora